Mouse anti rat cd31 biotin

Mesenteric angiogenesis was quantified using CD31‐labelled microvascular networks in rat mesenteric connective tissue windows according to a previous study.²⁶ At least four mesenteric windows (wedge‐shaped regions of connective tissue bordered by the intestinal wall and ileal blood vessel pairs) were dissected free in each rat, washed in PBS, dried on gelatin slides and fixed in 100% MeOH (−20℃ for 30 min). The slides were incubated overnight at 4℃ with the primary antibody mouse anti‐rat CD31‐biotin [1:200; AbD Serotec, Oxford, UK]. Secondary antibodies [CY2‐conjugated streptavidin, 1:1000; Jackson ImmunoResearch, West Grove, PA, USA] were then applied for 1 h at room temperature. At least four sets of data were obtained for each mesenteric window. (100×)‐magnification immunofluorescent images were assessed using an upright fluorescent microscope (AX80, Olympus, Japan) with a charge‐couple device (QICAM, High‐performance IEEE 1394 FireWireTM Digital CCD Camera, Q IMAGING, BD, Canada) and thresholded using Image J software (available for download from the National Institutes of Health (NIH, http://rsb.info.nih.gov/ij/). The vascular area was measured automatically using the histogram function.

Angiogenesis was quantified using CD31-labelled microvascular networks in rat mesenteric connective tissue windows according to a previous study with minor modifications [53 (link)]. From each rat, at least four mesenteric windows (wedge-shaped regions of connective tissue bordered by the intestinal wall and ileal blood vessel pairs) were dissected free and fixed in 100% MeOH (−20 °C for 30 min). Primary antibody mouse anti-rat CD31-biotin (1:200; AbD Serotec, Oxford, UK) and secondary antibody (CY2-conjugated streptavidin, 1:1000; Jackson ImmunoResearch, West Grove, PA, USA) were applied. At least four sets of data were obtained for each mesenteric window. Magnified immunofluorescent images (100×) were assessed using an upright fluorescent microscope (AX80, Olympus, Tokyo, Japan) with a charge-couple device (QICAM, High-performance IEEE 1394 FireWireTM Digital CCD Camera, Q IMAGING, BD, Canada) and thresholded by Image J software (available for download from the National Institutes of Health [54 ] as previously described [55 (link)].

Mesenteric angiogenesis was quantified by CD31-labelled microvascular networks in rat mesenteric connective tissue windows according to the previous study [29 (link)]. From each rat, at least four mesenteric windows (wedge-shaped regions of connective tissue surrounded by the intestinal wall and the ileal blood vessel pairs) were dissected free, washed in PBS, dried on gelatin slides, and fixed in 100% MeOH (−20 °C for 30 min). Slides were then incubated overnight at 4 °C with the primary antibody mouse anti-rat CD31-biotin (AbD Serotec, Oxford, UK). Then, a secondary antibody (CY2-conjugated streptavidin; Jackson ImmunoResearch, West Grove, PA, USA) was applied for 1 h at room temperature. At least four sets of data were obtained for each mesenteric window. Immunofluorescent images at magnification ×100 were assessed using an upright fluorescent microscope (AX80, Olympus, Tokyo, Japan) and thresholded by ImageJ software (ImageJ, Available online: https://imagej.nih.gov/ij/ (accessed on 6 June 2018). The vascular area was measured with the histogram function.